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1.
Article in Chinese | WPRIM | ID: wpr-940596

ABSTRACT

ObjectiveTo explore the structural characteristics and functional differences of intestinal flora in patients with type 2 diabetes mellitus (T2DM) of dampness heat trapping spleen(DHTS) syndrome and Qi-Yin deficiency(QYD) syndrome. MethodFrom June 2018 to January 2020,62 T2DM patients with DHTS syndrome and 60 with QYD syndrome were selected from Nanjing Hospital of Chinese Medicine Affiliated to Nanjing University of Chinese Medicine. Serum and fecal samples were collected to compare body mass index(BMI),glucose and lipid metabolism,fasting insulin (FINS) and fasting C-peptide (FCP) levels,and homeostasis model assessment of insulin resistance(HOMA-IR) of the two syndrome types. Fecal samples were extracted for DNA database construction,and 16S rDNA high-throughput sequencing was used to analyze and compare the intestinal flora and metabolic pathways. Result① The BMI,fasting plasma glucose(FPG),2-hour postprandial blood glucose (2 h PBG),total cholesterol(TC),triglyceride(TG),low density lipoprotein(LDL),FINS,FCP,and HOMA-IR were higher in patients with DHTS syndrome than in patients with QYD syndrome,and the high density lipoprotein(HDL) of the former was lower than that of the latter,(P<0.05,P<0.01). ② In terms of species composition and differences,Bacteroidetes, Clostridia and Gammaproteobacteria were dominant at the class level,and the relative abundance of Clostridia,Mollicutes and Verrucomicrobiae in QYD syndrome group was higher than that in DHTS syndrome group. At the order level,Bacteroidales,Clostridiales and Enterobacteriales were mainly found. The relative abundance of Clostridiales,Erysipelotrichales and Verrucomicrobiales in QYD syndrome group was obviously higher than that in DHTS syndrome group,while Aeromonadales in the former was lower than that in the latter (P<0.05). At the family level,Bacteroidaceae,Prevotellaceae and Ruminococcaceae were predominant. The relative abundance of Ruminococcaceae,Porphyromonadaceae and Erysipelotrichaceae in QYD syndrome group was higher than that in DHTS syndrome group(P<0.05). At the genus level,Bacteroides,Prevotella and Parabacteroides were mainly found. The relative abundance of Parabacteroides,Butyrivibrio and Ruminiclostridium in QYD syndrome group was higher than that in DHTS syndrome group,while that of Klebsiella and Megasphaera in DHTS syndrome group was higher than that in QYD syndrome group(P<0.05). ③ Through Venn analysis of operational taxonomic units(OTU),it was found that there were 49 OTUs in patients with DHTS syndrome patients and 47 OTUs in QYD syndrome patients. ④ The results of OTU β diversity and α analysis showed that Shannon and Simpson indexes had statistical differences,while Ace and Chao indexes had no statistical differences. The intestinal microbial diversity of patients with QYD syndrome was higher than that of patients with DHTS syndrome(P<0.05). The analysis of similarities (ANOSIM) showed that the difference of β diversity between the two groups was significant(P<0.05). ⑤ Linear discriminant analysis Effect Size(LEfSe) results demonstrated that Klebsiella,Megasphaera and Aeromonadales could be selected as the key biomarkers for DHTS syndrome; 14 bacteria such as Ruminiclostridium,Burkholderiaceae,Lautropia,Butyrivibrio,Erysipelotrichales can be selected as the key biomarkers for QYD syndrome. ⑥Functional annotation and analysis showed that the DHTS syndrome involved 9 metabolic pathways,including arginine and proline metabolism,lipopolysaccharide biosynthesis,nicotinic acid and nicotinamide metabolism,while the QYD syndrome involved 10 metabolic pathways,including acarbose and valinomycin biosynthesis,glucagon signaling pathway and NOD-like receptor signaling pathway. ConclusionThere are obvious differences in intestinal flora and functions in T2DM patients of DHTS syndrome and QYD syndrome,which can be used as reference for traditional Chinese medicine (TCM) syndrome differentiation and the target of TCM treatment.

2.
China Pharmacy ; (12): 51-55, 2021.
Article in Chinese | WPRIM | ID: wpr-862265

ABSTRACT

OBJECTIVE:To study the effects of shikonin on autophagy and apoptosis of human colon cancer HCT 116 cells. METHODS:After treating HCT 116 cells for 48 h with shikonin at 0(blank control )10,20,40 μg/mL,MTT method was used to detect inhibitory rate of cell proliferation. Flow cytometry was used to detect cell apoptosis rate. RT-qPCR assay and Western blotting were respectively used to detect mRNA and protein expressions of microtubule associated protein light chain 3(LC3)and autophagy-related protein Beclin- 1 and p 62. RESULTS :Compared with blank control ,after treated with 10,20,40 μ g/mL shikonin for 48 h,proliferation inhibitory rate and apoptosis rate of HCT 116 cells were increased significantly (P<0.05 or P< 0.01). After treated with 10,20,40 μg/mL shikonin for 48 h,mRNA and protein expressions of LC 3,Beclin-1 and p 62 in HCT116 cells were increased to different extents ;except that mRNA expression of LC 3 was not increased significantly after treated with 10 μg/mL shikonin,the difference were statistically significant in other indexes ,compared with blank control (P<0.05 or P<0.01). CONCLUSIONS :Shikonin can induce the apoptosis of human colon cancer HCT 116 cells and activate its autophagy pathway.

3.
China Pharmacy ; (12): 1537-1542, 2021.
Article in Chinese | WPRIM | ID: wpr-881452

ABSTRACT

OBJECTIVE:To provide reference for the c onstruction of subject diagnosis and treatment scheme in drug clinical trials. METHODS :The subject diagnosis and treatment module was developed and implemented in our hospital on the basis of CTMS,and its effects were evaluated. RESULTS :A subject diagnosis and treatment module was established in CTMS of our hospital. Within one year from the launch of the module in the middle of October ,2019,the overall number of subjects in the group showed an increasing trend ,and the overall mean dropout rate of subjects was 0.16%. The data interface of CTMS system , hospital information system (HIS),laboratory information management system ,medical imaging information system had been established,so as to realize the synchronization of subject information (displaying subject identification in HIS system )and the interaction of diagnosis and treatment information and billing data (patients and subjects were charged separately ). Since the launch of the module ,the amount of data generated by the interface had been increasing ,and the number of departments producing the subject diagnosis and treatment business had been increasing month by month. Compared with subject diagnosis and treatment project based on HIS system ,the number of subject diagnosis and treatment business based on CTMS system was increased significantly(P<0.05). CONCLUSIONS :The subject diagnosis and treatment module based on CTMS improves the efficiency of subject diagnosis and treatment project implementation and financial settlement ,and realizes the efficient implementation of drug clinical trial projects in large general hospitals.

4.
Article in Chinese | WPRIM | ID: wpr-863689

ABSTRACT

This paper summarizes the relevant literatures at home and abroad in recent years, and concludes that the active constituents of Aconiti radix treating rheumatoid arthritis mainly by reducing the tumor necrosis factor, interleukin-1, -6, -8, vascular endothelial growth factor, nuclear factor B, transient receptor potential vanillic acid. Receptor type 1 expression and up-regulating the nuclear factor E2-related factor 2 levels.

5.
Protein & Cell ; (12): 866-880, 2020.
Article in English | WPRIM | ID: wpr-880881

ABSTRACT

For multicellular organisms, cell-cell communication is essential to numerous biological processes. Drawing upon the latest development of single-cell RNA-sequencing (scRNA-seq), high-resolution transcriptomic data have deepened our understanding of cellular phenotype heterogeneity and composition of complex tissues, which enables systematic cell-cell communication studies at a single-cell level. We first summarize a common workflow of cell-cell communication study using scRNA-seq data, which often includes data preparation, construction of communication networks, and result validation. Two common strategies taken to uncover cell-cell communications are reviewed, e.g., physically vicinal structure-based and ligand-receptor interaction-based one. To conclude, challenges and current applications of cell-cell communication studies at a single-cell resolution are discussed in details and future perspectives are proposed.


Subject(s)
Animals , Cell Communication , Humans , RNA-Seq , Single-Cell Analysis , Transcriptome
6.
Article in Chinese | WPRIM | ID: wpr-802281

ABSTRACT

Objective:To establish HPLC-UV fingerprints of Ilex pubescens pieces,and simultaneously determine two components in 46 batches of I. pubescens in pieces of I. pubescens saponin A1 and B1,in order to provide a reference for the quality standard of I. pubescens slices. Method:Methanol was used to extract the I. pubescens saponin samples,and the extracts were measured by HPLC-UV with the absorption wavelength at 210 nm. Kromasil C18 column (4.6 mm×250 mm,5 μm) was used for determining the extracts at a flow rate of 1.0 mL·min-1. The mobile phase condition was acetonitrile-0.1% phosphoric acid aqueous solution with gradient mode. The chromatographic fingerprint similarity evaluation system of traditional Chinese medicine (2012 edition) was used to analyze I. pubescens fingerprints. SPSS 20.0 software was used to cluster the peak area of common peaks. Principal component analysis was performed to reduce the dimension of common peaks. Result:There were great differences between the root and stem parts in I. pubescens fingerprints. The fingerprints of roots and stems of I. pubescens were established respectively,cluster results assorted the roots of I. pubescens into three categories andthe branches of I. pubescens into two categories. The integrity and difference of I. pubescens decoction pieces from different parts and places of origin were compared,and the principal component analysis was performed to screen out the common components that played a decisive role in fingerprint of I. pubescens pieces. And the common peaks were determined. The content of saponin A1 and saponin B1 in Radix I. pubescens were determined. Conclusion:The established I. pubescens fingerprints and content determination methods are simple and suitable. Cluster analysis and principal component analysis are used to screen out the key components of quality control of I. pubescens. The results can provide references for quality control of I. pubescens.

7.
China Pharmacy ; (12): 2782-2788, 2019.
Article in Chinese | WPRIM | ID: wpr-817521

ABSTRACT

OBJECTIVE: To study the effects of benzoyl aconitine (BAC) on autophagy and apoptosis of human lung cancer A549 cells, and to investigate its mechanism in anti-non-small cell lung cancer. METHODS: A549 cells were treated with different doses of BAC (10, 50, 100, 200, 400 μmol/L), and then cell morphology was obtained; the proliferation inhibition rate of the cell was determined by CCK-8 assay. The cells were divided into control group (without drug), BAC low-dose and high-dose groups (200, 400 μmol/L). After treated with relevant drugs, the apoptosis rate of cells was determined by flow cytometry. The gene and protein expression of apoptosis-related factors Bcl-2, Bax, Caspase-3 as well as autophagy-related factors Beclin1, LC3, P62 were determined by RT-PCR and Western blotting assay. RESULTS: After treated with different doses of BAC, the cells were shrunken and sparsely arranged; inhibitory rate of cell proliferation was increased significantly in BAC 100, 200, 400 μmol/L groups (P<0.05 or P<0.01). Results of flow cytometry showed that the apoptotic rates of cells were increased to different extents in BAC low-dose and high-dose groups after treated for 24 and 48 h, in a concentration and time-dependent manner. Compared with control group, mRNA and protein expression of Bcl-2 and P62 were decreased to different extents in BAC groups; mRNA expression of Bax, Caspase-3, Beclin1 and LC3 as well as protein expression of Bax, Active caspase-3, P62, Beclin1, LC3Ⅱ/Ⅰ were increased to different extent; there was statistical significance in mRNA expression of Caspase-3, and protein expression of Bcl-2, Active Caspase-3, Beclin1, LC3Ⅱ/Ⅰ and P62 in BAC low-dose group as well as all target mRNA and protein expression in BAC high-dose group (P<0.05 or P<0.01), in dose-dependent manner. CONCLUSIONS: BAC can inhibit the proliferation and promote the apoptosis of A549 cells, promote Beclin1, LC3(LC3Ⅱ/Ⅰ),Bax and Caspase-3 (Active Caspase-3) gene and their protein expression, but inhibit P62 and Bcl-2 gene and their protein expression. The mechanism may be related to BAC inducing apoptosis by promoting excessive autophagy of cells.

8.
Article in Chinese | WPRIM | ID: wpr-905533

ABSTRACT

Objective:To design and implement a low-power and portable transcranial direct current stimulator controlled by mobile phones. Methods:The constant current stimulation circuit was realized by a field effect transistor, which could output stable and adjustable low-intensity direct current, and the impedance detection circuit and the over-current protection circuit increased the effectiveness and safety of the stimulator. The control and real-time detection of the stimulation circuit was realized through a microcontroller, and the parameters' settings of the stimulator and the display and preservation of the actual stimulus information were realized through the Android software on the smartphone. Results:The output current strength and accuracy, maximum load, as well as the timing, device connection, stimulus information collection and display all achieved the expected goals. Conclusion:The design realized the mobile control of the stimulator, with portability, low cost and low power consumption, providing a new solution for wider applications.

9.
Acta Pharmaceutica Sinica ; (12): 1123-1131, 2019.
Article in Chinese | WPRIM | ID: wpr-780173

ABSTRACT

This study aimed to construct an intelligent fluorescent nanocarrier for tumor cell tracing. Doxorubicin (DOX) was used as a model drug, and the gene targeting siBcl-2 was loaded to achieve synergistic inhibition of tumor cells. Mesoporous silicon nanoparticles (MSN) were prepared by a sol-gel method, and acetaldehyde cystine (AC) and polyethyleneimine (PEI) were covalently modified. The prepared nanocarrier MSN-AC-PEI was uniformly dispersed, with a particle size of 235.53 nm and a potential of 14.63 mV. The carrier material MSN-AC-PEI could load siRNA with the mass ratio of 60∶1 (Wvectors∶WsiRNA) and protect siRNA from RNase I degradation. To simulate the microenvironment of tumor, DOX release in phosphate buffer (pH 5) including 10 mmol·L-1 glutathione (GSH) was investigated. The cumulative release rate of DOX at 120 h is 35 times that of the normal physiological environment, which lays the foundation for the intelligent release of DOX in tumor cells. The results of cell experiments showed that the carrier material MSN-AC-PEI had significant green fluorescence, and the traceability can be maintained for 24 h after taken up by MCF-7 cells. After 24 hours of administration of the nano drug delivery system MSN-AC-PEI@DOX/siBcl-2, the inhibition rate of tumor cell proliferation reached 40.91%, and the late apoptosis rate was 60.84%. The Western blot results showed that compared with free DOX and siBcl-2, the nano-delivery system MSN-AC-PEI@DOX/siBcl-2 can significantly reduce the expression of anti-apoptotic protein Bcl-2, thereby enhancing its anti-tumor ability.

10.
Article in Chinese | WPRIM | ID: wpr-851906

ABSTRACT

Objective To study the effects of ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1, saikosaponin a, and saikosaponin d in Ginseng Radix et Rhizoma-Bupleuri Radix (GRRBR) herb pair and its preparations by using quantitative analysis of multi-components by single-marker (QAMS). Methods On the basis of ginsenoside Rg1, the relative correction factors between the ginsenoside Rg1 and the other four saponins were established, and then the contents of the other four saponins were calculated. At the same time, the contents of the five components were determined by external standard method and compared with those evaluated by QAMS. The relative retention time was determined by different chromatographic columns. It could be considered that QAMS was feasible and accurate in the determination of saponins in GRRBR herb pair. Results A quantitative control method of five kinds of saponins was established, and the methodological results were good. The results showed that there was no significant difference in the contents of five kinds of saponins in GRRBR herb pair, Xiaochaihu Decoction, and Kaixin Jieyu Prescription between QAMS group and external standard method group. Conclusion QAMS is suitable for the determination of saponin in GRRBR herb pair, which can be used as a reference for the determination of its effective components and the establishment of compound quality control method.

11.
China Pharmacy ; (12): 1259-1263, 2018.
Article in Chinese | WPRIM | ID: wpr-704778

ABSTRACT

OBJECTIVE:To evaluate the pharmacoeconomic characteristics of dapagliflozin combined with metformin in the treatment of type 2 diabetes mellitus systematically. METHODS:Retrieved from Health Technology Assessment(HTA),Cochrane Library,PubMed,Embase,CNKI,Wanfang and CBM during database establishment to Jul. 2017,published pharmacoeconomics literatures about dapagliflozin combined with metformin were collected,using"sodium-glucose-transporter-2 inhibitors""SGLT2 inhibitor""metformin""dapagliflozin""cost""benefit""utility""effectiveness""pharmacoeconomic""economic"as English retrieval words and"SGLT2 inhibitor""dage liejing""metformin""cost""benefit""utility""effectiveness"as Chinese retrieval words. Outcome indexes included incremental cost,incremental effect,cost-effectiveness ratio and incremental cost-effectiveness ratio(ICER).The results of the economic research in the included literatures were evaluated systematically. RESULTS:Totally of 4 literatures were included,and all of them were cost-effectiveness analysis. Dapagliflozin was more cost-effective than sulfonylurea because the ICER of dapagliflozin were €2 709/QALY,€10 494/QALY,€7 939/QALY,€5 433/QALY,€4 767/QALY and €6 094/QALY in the UK,Greece,Denmark,Finland,Norway and Sweden,respectively,which were all lower than willingness-to-pay threshold. Dapagliflozin was more cost-effective than DPP-4 inhibitor,and the ICER were €7 200/QALY and €15 120/QALY in the UK and Greece,respectively,which were all lower than willingness-to-pay threshold. CONCLUSIONS:Current economic research shows compared with sulfonylurea and DDP-4 inhibitor,dapagliflozin is a cost-effective treatment alternative for patients with T2DM whose metformin regimen does not provide sufficient glycemic control.

12.
Article in Chinese | WPRIM | ID: wpr-662086

ABSTRACT

Objective To design a medical cadre management system to improve cadre management in flexibility,rationality and openness.Methods The system was developed based on the traditional information management system,which used J2EE SSM framework to execute data management and logical operation and applied Wechat official account to implement cross-platform operation.Results The system facilitated cadre training and evaluation greatly.Conclusion The system contributes to collecting information on cadre evaluation and improving cadre training,and has practical values.

13.
Article in Chinese | WPRIM | ID: wpr-659359

ABSTRACT

Objective To design a medical cadre management system to improve cadre management in flexibility,rationality and openness.Methods The system was developed based on the traditional information management system,which used J2EE SSM framework to execute data management and logical operation and applied Wechat official account to implement cross-platform operation.Results The system facilitated cadre training and evaluation greatly.Conclusion The system contributes to collecting information on cadre evaluation and improving cadre training,and has practical values.

14.
Article in Chinese | WPRIM | ID: wpr-256085

ABSTRACT

Screening out the safety-related substances and establishing the corresponding standard has been a key research issue to improve the safety of traditional Chinese medicine injections(TCMIs). 5-HMF which widely exists in sugar-containing TCMIs has long been considered as an important safety-related substance. In this review, we summarizes the research progress on the toxicology of 5-HMF as well as the content and standards of 5-HMF in TCMIs.Therein, both literature summary and analysis results indicate that there are lack of toxicology researches of 5-HMF and its metabolites in TCMIs, although the potential toxicity of 5-HMF and its metabolites has been reported. Moreover, the content of 5-HMF largely varies from TCMIs to TCMIs, and even in the same TCMIs from different factories. To ensure the clinical efficacy of TCMIs, it urgent to carry out the study of the toxicology of 5-HMF in TCMIs comprehensively and systematically, so as to set up a relatively uniform standard as well as to develop process quality control method.

15.
Article in Chinese | WPRIM | ID: wpr-491916

ABSTRACT

The objective of this paper is to discover new potent inhibitors against EphB4 for cancer therapy via computer-aided drug design strategies including building 3D-QSAR models,virtual screening and molecular doc-king means.The first step is to generate pharmacophore models based on Catalyst/HypoGen algorithm.The best model,Hypo1,has the highest Correl value (0.96),the lowest RMS value (0.89),the closest total cost (101.26) to fixed cost (89.20),and the best Δcost (89.14).Subsequently,Hypo1 was subjected to test set validation and Fischer′s randomization verification and then was used as a 3D query to screen database.In order to further nar-row the number of hits,drug-likeness screening and molecular docking techniques were applied.Finally,23 novel molecules with diverse scaffolds were selected as possible candidates against EphB4 for further studies based on predicted activity analysis,docking scores,and binding modes analysis methods.

16.
Article in Chinese | WPRIM | ID: wpr-236059

ABSTRACT

To establish a method for the simultaneous determination of phloridzin, 3-hydroxy phloridzin and quercitrin in leaves of Malus halliana by ultrasonic-assisted ionic liquid coupled with RP-HPLC. An Agilent TC-C₁₈ (4.6 mm×250 mm, 5 μm) column was used, with the mobile phase of acetonitrile and 1% phosphoric acid-water (20∶80) by gradient elution at the detection wavelength of 270 nm. The flow rate was 0.8 mL•min⁻¹, and chromatographic column temperature was controlled at the room temperature. Under the optimized conditions, the linear ranges for phloridzin, 3-hydroxy phloridzin and quercitrin were 0.9-112.5 μg (r = 0.999 6), 0.093 2-11.65 μg (r = 0.999 1) and 0.097 2-12.15 μg (r = 0.999 8), respectively. The average recoveries of the three constituents were 99.35%, 98.80% and 98.19%, respectively. The method was environmental friendly, rapid, accurate and highly reproducible, and so suitable for the quantitative analysis of phloridzin, 3-hydroxy phloridzin and quercitrin in leaves of M. halliana.

17.
Article in Chinese | WPRIM | ID: wpr-470114

ABSTRACT

Objective To discuss the influence of individual cognitive on medical personnel blood-borne occupational exposure protection action from the angle of behavior operation.Methods Medical staff of 14 hospitals in zunyi were investigated by questionnaire designed based on the theory of health belief model,and analyzed the data by structural equation model.Results Sample data and the assumption model was ideal,the blood-borne occupational exposure protective behavior of medical staff could be explained variance of 87% by susceptibility,severity,behavioral benefit and barrier cognition.The order of influencing factors from high to low were behavioral benefit,severity,behavioral barrier and susceptibility to cognition,and path coefficients were 0.39,0.27,-0.21,0.03.Conclusions Susceptibility,severity and behavioral benefit cognition have positive effection on protective behavior,the behavioral benefit cognition have more influence on blood-borne occupational exposure protective behavior of medical staff,and behavioral barrier cognition have negative effection,the results of health belief model can explain blood-borne occupational exposure protective behavior of medical staff better.

18.
Article in Chinese | WPRIM | ID: wpr-459279

ABSTRACT

Objective To study the genotypes of human papillomavirus (HPV)infection in female anus and anal canal condylo-ma acuminata(CA)tissues and their clinical significance.Methods 23 kinds of HPV-DNA were extracted from the paraffin-embed-ded anus and anal canal tissue samples in 140 cases of female CA and detected by using PCR combined with the gene-chips tech-nique.Furthermore the related clinical pathological data of the patients were analyzed.Results Among 140 female anus and anal ca-nal CA tissue samples,103 cases were HPV positive and the total HPV infection rate was 73.57%(103/140).Among them,68 ca-ses were single type HPV infection,the positive detection rate was 48.57%(68/140)and 35 cases were multiple types HPV infec-tion,the positive detection rate was 25.00% (35/140).In single type HPV infection,34 cases were HPV11 and the positive detec-tion rate was 24.29% (34/140),HPV11 was the main infection type,followed by HPV 6 in 27 cases,its positive detection rate was 19.29%(27/140).In the multiple types HPV infection,13 cases were HPV 6 + 11,accounting for 37.14% (13/35 )of multiple types infection,followed by HPV11 +18 in 3 cases and HPV 6+11+16 in 3 cases,each accounting for 8.57%(3/35)of the multi-ple types infection.Conclusion HPV 6,11 ,6+11,11 +18 and 6+11+16 are the main infection genotypes in female anus and anal canal CA.PCR combined with the gene-chips technique is a diagnostic method more suitable for clinical development of HPV geno-typing detection,which has high sensitivity and good specificity and is especially suitable for the molecular epidemiology study of HPV infection.

19.
Article in English | WPRIM | ID: wpr-347190

ABSTRACT

Chinese medicine (CM) clinical efficacy evaluation research involves the longitudinal multivariate measurement which means that patients are measured repeatedly and each patient is measured by several indicators on each fixed cross-section. Although each indicator can be evaluated separately with a longitudinal linear mixed model, it is important to consider all the endpoints together especially when researchers pay special attention to the change of the conjoint efficacy for several indicators in one patient. In this article, we introduce a latent variable linear mixed model to the CM conjoint efficacy evaluation and discuss why and how to analyze the longitudinal multivariate endpoint data in the clinical CM efficacy evaluation research. It may lead to the new insight of using such methodology in the field of conjoint efficacy evaluating of CM study. And with the definition of syndrome and symptom in the CM theory, the applied discussion brings the insight of CM syndrome evaluating in future. We illustrate this methodology using an example of CM efficacy evaluation from an ischemic stroke research.


Subject(s)
Biomedical Research , Methods , Data Interpretation, Statistical , Endpoint Determination , Methods , Humans , Linear Models , Medicine, Chinese Traditional , Methods , Multivariate Analysis , Research Design , Stroke , Drug Therapy , Epidemiology , Treatment Outcome
20.
Chinese Medical Journal ; (24): 2523-2529, 2013.
Article in English | WPRIM | ID: wpr-322168

ABSTRACT

<p><b>BACKGROUND</b>In vitro chondrocyte expansion is a major challenge in cell-based therapy for human articular cartilage repair. Classical culture conditions usually use animal serum as a medium supplement, which raises a number of undesirable questions. In the present study, two kinds of defined, serum-free media were developed to expand chondrocytes in monolayer culture for the purpose of cartilage tissue engineering.</p><p><b>METHODS</b>Bovine chondrocytes were expanded in serum-free media supplemented with fibroblast growth factor-2 and platelet-derived growth factor or fibroblast growth factor-2 and insulin-like growth factor. Expansion culture in a conventional 10% fetal bovine serum (FBS) medium served as control. Fibronectin coating was used to help cell adhesion in serum-free medium. Next, in vitro three-dimensional pellet culture was used to evaluate the chondrocyte capacity. Cell pellets were expanded in different media to re-express the differentiated phenotype (re-differentiation) and to form cartilaginous tissue. The pellets were assessed by glycosaminoglycans contents, collagen II, collagen I and collagen X immunohistological staining.</p><p><b>RESULTS</b>Chondrocytes cultured in serum-free media showed no proliferation difference than cells grown with 10% FBS medium. In addition, chondrocytes expanded in both serum-free media expressed more differentiated phenotypes at the end of monolayer culture, as indicated by higher gene expression ratios of collagen type II to collagen type I. Pellets derived from chondrocytes cultured in both serum-free media displayed comparable chondrogenic capacities to pellets from cells expanded in 10% FBS medium.</p><p><b>CONCLUSION</b>These findings provide alternative culture approaches for chondrocytes in vitro expansion, which may benefit the clinical use of autologous chondrocytes implantation.</p>


Subject(s)
Animals , Cartilage, Articular , Cell Biology , Cattle , Cell Dedifferentiation , Cells, Cultured , Chondrocytes , Cell Biology , Physiology , Culture Media, Serum-Free , Fibronectins , Pharmacology , Real-Time Polymerase Chain Reaction , SOX9 Transcription Factor , Genetics
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